The Influence Of Rukangfang Decoction On Serum Cytokines (IL-2, TNF-α) In Rat Mammary Gland Hyperplasia Model

Objective: Mammary gland hyperplasia is a hyperplastic neuopathy which is not inflammation nor tumor. It is a kind of disease generalterm, whose pathological changes including mamary gland alveoluses hyperplasia, mamary gland ductal epithelial cell hyperplasia and fibrous connective tissue hyperplasia. Women at child-bearing age are easy to have MGH. Pathogenetic peak age of MGH is 35～45 years old. Its incidence rate accounts for 75% of all the breast diseases. The crowd with breast cancer have 2～3 times incidence than the common. So, active treatment in MGH has much important significance in preventing breast cancer, and also can improve quality of life of female patients. To existenting data, modern medicine is still lack of satisfactory therapeutic measure MGH. Its endocrine therapy has much adverse reaction, and long-time treatment may interfere the endocrine in balance. Therefore, it is not suitable as routine treatment. Our study based on that Rukangfang decoction effectively therapy on MGH in clinical practice, we established rat models of MGH. Compaired chinese patent medicine Rupixiao, TAM with Rukangfang decoction, we observed curative effect of Rukangfang decoction and revealed partial pathogenesis of MGH in immunology, through analyzing level of IL-2, TNF-αin serum and mammary tissue pathological changes with MGH, we investigate mechanism of Rukangfang therapy on MGH. It will enrich and develop practitioner of Chinese traditional medicine.Method: 70 healthy Wistar female rats in cleanness grade,75-85 days, 220±20g, after raising by normal diet for a week, were divided into 6 groups randomly: Normal group, model group, Rupixiao control group, TAM control group, low dose of Rukangfang group (Rukangfang I group), high dose of Rukangfang group (RukangfangⅡgroup). The rats in normal group were treated with the injection of normal saline on the right lower limb, in dose of 0.2ml/d, once a day for 30 days. In other group, those rats were injected estradiol benzoate, in dose of 0.5mg/kg, once a day for 25 days, then injected progesterone, in dose of 5mg/kg, once a days for 5 days. From the thirty-first day, we extracted separately 5 rats from the normal group and the model group by using the random number table, then measured the second mammary diameter and cut the third mammary. Results: observing pathological sections the by optical microscope, it showed that mammary gland ducts and mamary gland alveoluses were proliferated obviously compaired with normal group. The results indicated that we established the rat MGH model successfully. Atfter modeling successfully, each group adopted group therapy. Each group was drench in the dosage of 10ml/kg every day with Rupixiao suspension, TAM suspension, low dose of Rukangfang decoction and high low of Rukangfang decoction, and with distilled water in normal and model group all for 30 days. Fasting 24 hours after last drench, measured the second mammary diameter with sliding caliper, collected blood by decollation, placed them respectively, detected the serum level of estradiol(E2), progestogen(P), prolactin(PRL), Interleukin-2 (IL-2) and Tunmor Necrosis Factor (TNF-α) by means of radioimmunology. Cut rats mammary quickly, slightly wiped with filter paper, fixed mammary in 4% polyoxymethylene, paraffin emdedded, slice in HE dyeing, observed the mammary histomorphology.Results:1 changes of rat mammary diameterCompaired with normal group, the mammary diameters in model group were bigger, and there were significant difference between two groups (P<0.01). After therapy, mammary diameters in Rupixiao group and TAM group were than that in model group, the difference was of statistical significance (P<0.05). The mammary diameters in Rukangfang I group and RukangfangⅡgroup, compaired with model group, were much shorter (P<0.01).2 Changes of IL-2 and TNF-αin serumCompaired with normal group, the level of serum IL-2 in model group was decreased, and there were significant difference between two groups (P<0.05). After therapy, there were an increasing tendency in Rupixiao group , TAM group and RukangfangⅠgroup, the difference was of statistical significance (P<0.05). The level of serum IL-2 in RukangfangⅡgroup was remarkably increased, compaired with model group, and there were significant difference (P<0.01).Compaired with normal group, the level of serum TNF-αin model group was increased, and there were significant difference between two groups (P<0.05). After therapy, there were an decreasing tendency in Rupixiao group, RukangfangⅠgroup and TAM group, compaired with model group, it was of no statistical significance (P>0.05). The level of serum TNF-αin RukangfangⅡgroup was remarkably decreased, there were significant difference (P<0.05).3 Changes of genital hormone in serumCompaired with normal group, the contents of serum E2 in model group were remarkably increased, and there were significant difference between two groups (P<0.01). After therapy, contents in Rupixiao group, TAM group and Rukangfang I group were decreased, and the difference was of statistical significance (P<0.05). The contents in RukangfangⅡgroup, compaired with model group, were remarkably decreased, and there were significant difference (P<0.01).Compaired with normal group, the contents of serum P in model group were remarkably decreased, and there were significant difference between two groups (P<0.01). After therapy, contents in Rupixiao group and Rukangfang I group all had an increasing tendency, but it was of no statistical significance (P>0.05). Its contents TAM group group were increased, cmpaired with model group, and there were significant difference (P<0.05). Its contents in RukangfangⅡgroup were remarkably increased, cmpaired with model group, and there were significant difference (P<0.01).Compaired with normal group, the contents of serum PRL in model group were remarkably increased, and there were significant difference between two groups (P<0.01). After therapy, contents in Rupixiao group, TAM group and Rukangfang I group all had a decreasing tendency, there were significant difference (P<0.05). Its contents in RukangfangⅡgroup were decreased, compaired with model group, and there were significant difference (P<0.01).Conclusion:1 The mammary diameters in model group were bigger than that in normal group. Pathological pictures show that mammary gland tissues appeared diffuse hyperplasy, acinar cells and lobules remarkably increased, acinar cavity remarkably expand. It accords with clinical standard and indicates that we established the rat MGH model successfully.2 Compaired with normal group, the serum level of IL-2 decreased in model group and the serum level of TNF-αincreased, it indicates that MGH rats exist disturbance of immune system and disturbance of immune system was the important factors of MGH pathogenesis. The high dose of Rukangfang group can increase the serum level of IL-2 and decreased the serum level of TNF-α, it can enhance immunity and promote the recovery of mammary gland tissue in the rat MGH model. The experiment reveal a part mechanism of Rukangfang therapy on MGH in immunology. This experiment also indicates that therapeutic effects of the high dose of Rukangfang decoction is superior to that of the low dose of Rukngfang decoction.3 Rukangfang decoction can adjust genital hormones of E2, P, PRL and immunity system, improve endocrine secretion, and achieve curing MGH. The high dose of Rukangfang decoction has significant efficiency than Rupixiao, TAM and the low dose of Rukangfang decoction.